'True' single-molecule molecule observations by fluorescence correlation spectroscopy and two-color fluorescence cross-correlation spectroscopy
'True' single-molecule molecule observations by fluorescence correlation spectroscopy and two-color fluorescence cross-correlation spectroscopy
复制标题
DOI：
10.1016/j.yexmp.2006.12.002
复制
发表时间：
2007-04-01
影响因子：
3.6
通讯作者：
Foeldes-Papp, Zeno
中科院分区：
文献类型：
--
作者：
Foeldes-Papp, Zeno
荧光相关光谱(FCS)和双色荧光互相关光谱(FCCS)是一种测量由于紧密聚焦的激光和共焦光学器件引起的荧光分子通过肥摩尔计检测体积时被检测光的波动。来自单个分子的荧光很容易与与溶剂的肥料有关的轻微背景区分开来。当溶液浓度约为1 nm时，探针体积中存在分析物分子的概率小于1。尽管收集了单个分子的荧光，但数据是通过自相关或双色互相关函数进行分析的，这些函数是数千个分子的平均值。不能获得单分子的性质。我一直在研究与非常稀薄的解决方案相关的问题和机会。共焦探针体积中的分子最有可能是刚刚扩散、转身和扩散回来的分子，即重新进入。首次提出了新的有意义时间理论的一些理论结果，该理论可以在不固定或流体动力学聚焦的情况下，在较长的时间内只研究一个单分子。重入也可称为单个分子的重现或相遇是重要的，因为在测量时间内，它们会引起波动现象，如分子数波动。同样，已经制定了四个标准，可以用来验证在实验期间激光探测器体积中只有一个“自我相同”的分子：(Foldes-Papp，Z.，2006)。用荧光涨落光谱测量溶液中的单个分子意味着什么。实验摩尔。帕托尔。80(3)209-218)。(C)2006 Elsevier Inc.保留所有权利。
Fluorescence correlation spectroscopy (FCS) and two-color fluorescence cross-correlation spectroscopy (FCCS) are a measure of fluctuations of detected light as a fluorescence molecule diffuses through a fermoliter detection volume caused by a tightly focused laser and confocal optics. Fluorescence from a single molecule can easily be distinguished from the slight background associated with a fermoliter of solvent. At a solution concentration of about 1 nM, the probability that there is an analyte molecule in the probe volume is less than one. Although fluorescence from individual molecules is collected, the data are analyzed by autocorrelation or two-color cross-correlation functions that are the average of thousands of molecules. Properties of single molecules are not obtained. I have been working on problems and opportunities associated with very dilute solutions. The molecule in the confocal probe volume is most probably the molecule that just diffused out, turned around, and diffused back in, i.e., reentered. For the first time, some theoretical results of the novel theory of the meaningful time are presented that enable study of just one single molecule over extended periods of times without immobilization or hydrodynamic focusing. Reentries that may also be called reoccurrences or encounters of a single molecule are significant because during measurement times they give rise to fluctuation phenomena such as molecule number fluctuations. Likewise, four criteria have been developed that can be used to verify that there is only one "selfsame" molecule in the laser probe volume during the experiment: (Foldes-Papp, Z., 2006. What it means to measure a single molecule in a solution by fluorescence fluctuation spectroscopy. Exp. Mol. Pathol. 80 (3) 209-218). (C) 2006 Elsevier Inc. All rights reserved.